Expression of fungal acetyl xylan esterase in Arabidopsis thaliana improves saccharification of stem lignocellulose

Prashant Mohan-Anupama Pawar , Marta Derba-Maceluch, Sun-Li Chong, Leonardo D. Gomez, Eva Miedes, Alicja Banasiak, Christine Ratke, Cyril Gaertner, Gregory Mouille, Simon J. McQueen-Mason, Antonio Molina, Anita Sellstedt, Tiina Maija Tenkanen, Ewa J. Mellerowicz

Forskningsoutput: TidskriftsbidragArtikelVetenskapligPeer review

Sammanfattning

Cell wall hemicelluloses and pectins are O-acetylated at specific positions, but the significance of these substitutions is poorly understood. Using a transgenic approach, we investigated how reducing the extent of O-acetylation in xylan affects cell wall chemistry, plant performance and the recalcitrance of lignocellulose to saccharification. The Aspergillus niger acetyl xylan esterase AnAXE1 was expressed in Arabidopsis under the control of either the constitutively expressed 35S CAMV promoter or a woody-tissue-specific GT43B aspen promoter, and the protein was targeted to the apoplast by its native signal peptide, resulting in elevated acetyl esterase activity in soluble and wall-bound protein extracts and reduced xylan acetylation. No significant alterations in cell wall composition were observed in the transgenic lines, but their xylans were more easily digested by a beta-1,4-endoxylanase, and more readily extracted by hot water, acids or alkali. Enzymatic saccharification of lignocellulose after hot water and alkali pretreatments produced up to 20% more reducing sugars in several lines. Fermentation by Trametes versicolor of tissue hydrolysates from the line with a 30% reduction in acetyl content yielded similar to 70% more ethanol compared with wild type. Plants expressing 35S: AnAXE1 and pGT43B:AnAXE1 developed normally and showed increased resistance to the biotrophic pathogen Hyaloperonospora arabidopsidis, probably due to constitutive activation of defence pathways. However, unintended changes in xyloglucan and pectin acetylation were only observed in 35S: AnAXE1-expressing plants. This study demonstrates that postsynthetic xylan deacetylation in woody tissues is a promising strategy for optimizing lignocellulosic biomass for biofuel production.
Originalspråkengelska
TidskriftPlant biotechnology journal
Volym14
Utgåva1
Sidor (från-till)387-397
Antal sidor11
ISSN1467-7644
DOI
StatusPublicerad - 2016
MoE-publikationstypA1 Tidskriftsartikel-refererad

Vetenskapsgrenar

  • 1183 Växtbiologi, mikrobiologi, virologi

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Mohan-Anupama Pawar , P., Derba-Maceluch, M., Chong, S-L., Gomez, L. D., Miedes, E., Banasiak, A., ... Mellerowicz, E. J. (2016). Expression of fungal acetyl xylan esterase in Arabidopsis thaliana improves saccharification of stem lignocellulose. Plant biotechnology journal, 14(1), 387-397. https://doi.org/10.1111/pbi.12393
Mohan-Anupama Pawar , Prashant ; Derba-Maceluch, Marta ; Chong, Sun-Li ; Gomez, Leonardo D. ; Miedes, Eva ; Banasiak, Alicja ; Ratke, Christine ; Gaertner, Cyril ; Mouille, Gregory ; McQueen-Mason, Simon J. ; Molina, Antonio ; Sellstedt, Anita ; Tenkanen, Tiina Maija ; Mellerowicz, Ewa J. / Expression of fungal acetyl xylan esterase in Arabidopsis thaliana improves saccharification of stem lignocellulose. I: Plant biotechnology journal. 2016 ; Vol. 14, Nr. 1. s. 387-397.
@article{6d9c8406a810445dbe3bce2f1c28f2fa,
title = "Expression of fungal acetyl xylan esterase in Arabidopsis thaliana improves saccharification of stem lignocellulose",
abstract = "Cell wall hemicelluloses and pectins are O-acetylated at specific positions, but the significance of these substitutions is poorly understood. Using a transgenic approach, we investigated how reducing the extent of O-acetylation in xylan affects cell wall chemistry, plant performance and the recalcitrance of lignocellulose to saccharification. The Aspergillus niger acetyl xylan esterase AnAXE1 was expressed in Arabidopsis under the control of either the constitutively expressed 35S CAMV promoter or a woody-tissue-specific GT43B aspen promoter, and the protein was targeted to the apoplast by its native signal peptide, resulting in elevated acetyl esterase activity in soluble and wall-bound protein extracts and reduced xylan acetylation. No significant alterations in cell wall composition were observed in the transgenic lines, but their xylans were more easily digested by a beta-1,4-endoxylanase, and more readily extracted by hot water, acids or alkali. Enzymatic saccharification of lignocellulose after hot water and alkali pretreatments produced up to 20{\%} more reducing sugars in several lines. Fermentation by Trametes versicolor of tissue hydrolysates from the line with a 30{\%} reduction in acetyl content yielded similar to 70{\%} more ethanol compared with wild type. Plants expressing 35S: AnAXE1 and pGT43B:AnAXE1 developed normally and showed increased resistance to the biotrophic pathogen Hyaloperonospora arabidopsidis, probably due to constitutive activation of defence pathways. However, unintended changes in xyloglucan and pectin acetylation were only observed in 35S: AnAXE1-expressing plants. This study demonstrates that postsynthetic xylan deacetylation in woody tissues is a promising strategy for optimizing lignocellulosic biomass for biofuel production.",
keywords = "1183 Plant biology, microbiology, virology, acetyl xylan esterase , biofuels , saccharification , O-acetylation , glucuronoxylan , secondary cell wall , WRKY60 TRANSCRIPTION FACTORS , CELL WALL POLYSACCHARIDES , O-ACETYLATION , SACCHAROMYCES-CEREVISIAE , RELATIVE QUANTIFICATION , ENZYMATIC-HYDROLYSIS , INCREASED RESISTANCE , XYLOSE FERMENTATION , ASPERGILLUS-NIGER , BOTRYTIS-CINEREA",
author = "{Mohan-Anupama Pawar}, Prashant and Marta Derba-Maceluch and Sun-Li Chong and Gomez, {Leonardo D.} and Eva Miedes and Alicja Banasiak and Christine Ratke and Cyril Gaertner and Gregory Mouille and McQueen-Mason, {Simon J.} and Antonio Molina and Anita Sellstedt and Tenkanen, {Tiina Maija} and Mellerowicz, {Ewa J.}",
year = "2016",
doi = "10.1111/pbi.12393",
language = "English",
volume = "14",
pages = "387--397",
journal = "Plant biotechnology journal",
issn = "1467-7644",
publisher = "Wiley",
number = "1",

}

Mohan-Anupama Pawar , P, Derba-Maceluch, M, Chong, S-L, Gomez, LD, Miedes, E, Banasiak, A, Ratke, C, Gaertner, C, Mouille, G, McQueen-Mason, SJ, Molina, A, Sellstedt, A, Tenkanen, TM & Mellerowicz, EJ 2016, 'Expression of fungal acetyl xylan esterase in Arabidopsis thaliana improves saccharification of stem lignocellulose', Plant biotechnology journal, vol. 14, nr. 1, s. 387-397. https://doi.org/10.1111/pbi.12393

Expression of fungal acetyl xylan esterase in Arabidopsis thaliana improves saccharification of stem lignocellulose. / Mohan-Anupama Pawar , Prashant; Derba-Maceluch, Marta; Chong, Sun-Li; Gomez, Leonardo D.; Miedes, Eva; Banasiak, Alicja; Ratke, Christine; Gaertner, Cyril; Mouille, Gregory; McQueen-Mason, Simon J.; Molina, Antonio; Sellstedt, Anita; Tenkanen, Tiina Maija; Mellerowicz, Ewa J.

I: Plant biotechnology journal, Vol. 14, Nr. 1, 2016, s. 387-397.

Forskningsoutput: TidskriftsbidragArtikelVetenskapligPeer review

TY - JOUR

T1 - Expression of fungal acetyl xylan esterase in Arabidopsis thaliana improves saccharification of stem lignocellulose

AU - Mohan-Anupama Pawar , Prashant

AU - Derba-Maceluch, Marta

AU - Chong, Sun-Li

AU - Gomez, Leonardo D.

AU - Miedes, Eva

AU - Banasiak, Alicja

AU - Ratke, Christine

AU - Gaertner, Cyril

AU - Mouille, Gregory

AU - McQueen-Mason, Simon J.

AU - Molina, Antonio

AU - Sellstedt, Anita

AU - Tenkanen, Tiina Maija

AU - Mellerowicz, Ewa J.

PY - 2016

Y1 - 2016

N2 - Cell wall hemicelluloses and pectins are O-acetylated at specific positions, but the significance of these substitutions is poorly understood. Using a transgenic approach, we investigated how reducing the extent of O-acetylation in xylan affects cell wall chemistry, plant performance and the recalcitrance of lignocellulose to saccharification. The Aspergillus niger acetyl xylan esterase AnAXE1 was expressed in Arabidopsis under the control of either the constitutively expressed 35S CAMV promoter or a woody-tissue-specific GT43B aspen promoter, and the protein was targeted to the apoplast by its native signal peptide, resulting in elevated acetyl esterase activity in soluble and wall-bound protein extracts and reduced xylan acetylation. No significant alterations in cell wall composition were observed in the transgenic lines, but their xylans were more easily digested by a beta-1,4-endoxylanase, and more readily extracted by hot water, acids or alkali. Enzymatic saccharification of lignocellulose after hot water and alkali pretreatments produced up to 20% more reducing sugars in several lines. Fermentation by Trametes versicolor of tissue hydrolysates from the line with a 30% reduction in acetyl content yielded similar to 70% more ethanol compared with wild type. Plants expressing 35S: AnAXE1 and pGT43B:AnAXE1 developed normally and showed increased resistance to the biotrophic pathogen Hyaloperonospora arabidopsidis, probably due to constitutive activation of defence pathways. However, unintended changes in xyloglucan and pectin acetylation were only observed in 35S: AnAXE1-expressing plants. This study demonstrates that postsynthetic xylan deacetylation in woody tissues is a promising strategy for optimizing lignocellulosic biomass for biofuel production.

AB - Cell wall hemicelluloses and pectins are O-acetylated at specific positions, but the significance of these substitutions is poorly understood. Using a transgenic approach, we investigated how reducing the extent of O-acetylation in xylan affects cell wall chemistry, plant performance and the recalcitrance of lignocellulose to saccharification. The Aspergillus niger acetyl xylan esterase AnAXE1 was expressed in Arabidopsis under the control of either the constitutively expressed 35S CAMV promoter or a woody-tissue-specific GT43B aspen promoter, and the protein was targeted to the apoplast by its native signal peptide, resulting in elevated acetyl esterase activity in soluble and wall-bound protein extracts and reduced xylan acetylation. No significant alterations in cell wall composition were observed in the transgenic lines, but their xylans were more easily digested by a beta-1,4-endoxylanase, and more readily extracted by hot water, acids or alkali. Enzymatic saccharification of lignocellulose after hot water and alkali pretreatments produced up to 20% more reducing sugars in several lines. Fermentation by Trametes versicolor of tissue hydrolysates from the line with a 30% reduction in acetyl content yielded similar to 70% more ethanol compared with wild type. Plants expressing 35S: AnAXE1 and pGT43B:AnAXE1 developed normally and showed increased resistance to the biotrophic pathogen Hyaloperonospora arabidopsidis, probably due to constitutive activation of defence pathways. However, unintended changes in xyloglucan and pectin acetylation were only observed in 35S: AnAXE1-expressing plants. This study demonstrates that postsynthetic xylan deacetylation in woody tissues is a promising strategy for optimizing lignocellulosic biomass for biofuel production.

KW - 1183 Plant biology, microbiology, virology

KW - acetyl xylan esterase

KW - biofuels

KW - saccharification

KW - O-acetylation

KW - glucuronoxylan

KW - secondary cell wall

KW - WRKY60 TRANSCRIPTION FACTORS

KW - CELL WALL POLYSACCHARIDES

KW - O-ACETYLATION

KW - SACCHAROMYCES-CEREVISIAE

KW - RELATIVE QUANTIFICATION

KW - ENZYMATIC-HYDROLYSIS

KW - INCREASED RESISTANCE

KW - XYLOSE FERMENTATION

KW - ASPERGILLUS-NIGER

KW - BOTRYTIS-CINEREA

U2 - 10.1111/pbi.12393

DO - 10.1111/pbi.12393

M3 - Article

VL - 14

SP - 387

EP - 397

JO - Plant biotechnology journal

JF - Plant biotechnology journal

SN - 1467-7644

IS - 1

ER -